%0 Journal Article
%T Analysis of mouse liver membrane proteins using multidimensional ion exchange chromatography and tandem mass spectrometry<br>多维离子交换色谱分离和串联质谱鉴定在小鼠肝脏膜蛋白质组分析中的应用
%A WANG Zhuowei
%A PENG Fuli
%A WANG Yuan
%A TONG Wei
%A REN Yan
%A XU Ningzhi
%A LIU Siqi
%A <br>王灼维
%A 彭福利
%A 王媛
%A 童维
%A 任艳
%A 徐宁志
%A 刘斯奇
%J 色谱
%D 2010
%I 
%X The analysis of membrane proteins is still a technical obstacle in proteomic investigation. A fundamental question is how to allow the hydrophobic proteins fully solubilizing in a proper solvent environment. We propose that the denatured membrane proteins in high denaturant solution are fully ionized and separated through ion exchange chromatography. The membrane proteins prepared from a mouse liver were dissolved in 4 mol/L urea, 20 mmol/L Tris-HCl buffer (pH 9.0), and loaded onto a tandem chromatography coupled with Q-Sepharose FF and Sephacryl S-200HR. With a linear NaCl gradient elution, the bound proteins were eluted and collected followed by sodium-dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) to further separate the eluted proteins. The protein bound on SDS-PAGE were excised and in-gel digested by trypsin, while the digested peptides were delivered to reversed-phase high performance liquid chromatography (HPLC) and ion-trap mass spectrometry for the peptide identifications. Of a total of 392 proteins identified, 306 were membrane proteins or membrane associated proteins reported by literature. Based on the calculation of hydrophobicity, the GRAVY (grand average of hydropathicity) scores of 83 proteins are over or equal to 0.00. Taking all the evidence, we have established an effective approach which is feasible in the investigation towards mouse liver membrane proteomics.
%K multidimensional ion-exchange chromatography
%K sodium-dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)
%K reversed-phase high performance liquid chromatography (RP-HPLC)
%K tandem mass spectrometry (MS/MS)
%K membrane protein
%K mouse liver<br>多维离子交换色谱
%K 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳
%K 反相高效液相色谱
%K 串联质谱
%K 膜蛋白质
%K 小鼠肝脏
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=6E709DC38FA1D09A4B578DD0906875B5B44D4D294832BB8E&cid=6579068328FE643F&jid=4D81E042D77AFEC6881D14759692069C&aid=5ADD7FBAEF6647E24DDA251329873DB3&yid=140ECF96957D60B2&vid=D3E34374A0D77D7F&iid=0B39A22176CE99FB&sid=EDA22B444205D04A&eid=B62E0EEFE746E568&journal_id=1000-8713&journal_name=色谱&referenced_num=0&reference_num=20